TY - JOUR T1 - Cervico-vaginal secretion cytokine profile: A non-invasive approach to study the endometrial receptivity in IVF cycles. JF - Am J Reprod Immunol Y1 - 2019 A1 - Zanotta, Nunzia A1 - Monasta, Lorenzo A1 - Skerk, Kristina A1 - Luppi, Stefania A1 - Martinelli, Monica A1 - Ricci, Giuseppe A1 - Comar, Manola AB -

PROBLEM: Cytokines have a significant role in the process of embryo implantation, trophoblast growth, and differentiation by modulating the immune and endocrine system. The aim of this study was to investigate the profile of a large set of cytokines in the cervico-vaginal washing of women undergoing IVF, to explore the association of these proteins with a good receptive endometrium.

METHOD OF STUDY: A cohort of 155 women scheduled for IVF cycle was recruited. All patients were asymptomatic for genitourinary infections and had been screened for chlamydia, mycoplasma, and other bacterial infections. All IVF subjects were treated according to standard clinical and laboratory protocols. A panel of 48 immune factors was analyzed on cervico-vaginal washing, using magnetic bead-based multiplex immunoassays (Bio-Plex, BIO-RAD Laboratories, Milano, Italy).

RESULTS: A total of 99 patients reached embryo transfer, of which 31 had a clinical pregnancy. A pattern of four pro-inflammatory immune molecules, IL-12p40, IFN-a, MIF, and MCP3 (P < 0.001), was found significantly up-regulated in the cervico-vaginal fluid of women with clinical pregnancy. A significantly increased expression of IL-9, Gro , and SDF-1 (P < 0.05) was observed in the presence of endometriosis, while high levels of IL-13 and L-15 were associated with ovulatory infertility factor (P < 0.05).

CONCLUSION: In this pilot study, we demonstrated that the expression of specific cytokines in the cervico-vaginal washing on the day of oocyte retrieval might have a positive correlation with the potential clinical pregnancy. Therefore, cervico-vaginal secretion cytokine profiling might be a new, non-invasive approach to study the endometrial receptivity in IVF management.

VL - 81 IS - 1 U1 - http://www.ncbi.nlm.nih.gov/pubmed/30475413?dopt=Abstract ER - TY - JOUR T1 - Thrombogenesis in Thrombophilic Pregnancy: Evaluation of Low-Molecular-Weight Heparin Prophylaxis. JF - Acta Haematol Y1 - 2017 A1 - Simeone, Roberto A1 - Giacomello, Roberta A1 - Bruno, Germano A1 - Parco, Sergio A1 - Maximova, Natalia A1 - Martinelli, Monica A1 - Zito, Gabriella A1 - Luppi, Stefania A1 - Cervi, Gina A1 - Ricci, Giuseppe KW - Adult KW - Anticoagulants KW - Case-Control Studies KW - Factor Xa Inhibitors KW - Female KW - Heparin, Low-Molecular-Weight KW - Humans KW - Nadroparin KW - Partial Thromboplastin Time KW - Peptide Fragments KW - Pilot Projects KW - Pregnancy KW - Pregnancy Complications, Hematologic KW - Prothrombin KW - Thrombophilia KW - Thrombosis AB -

The aim of this study is to investigate thrombogenesis and the hypercoagulable changes in pregnant women affected by thrombophilia who received low-molecular-weight heparin (LWMH) prophylaxis. We included 21 pregnant women affected by thrombophilia treated with LWMH and 20 nontreated normal pregnant women as the control group. The sample group of thrombophilic pregnant women included different conditions (factor V Leiden mutation, protein C deficiency, protein S deficiency, antiphospholipid antibodies syndrome, and combined defects). Three blood samples were collected during pregnancy (i.e., at 16, 20, and 24 weeks) and tested for activated partial thromboplastin time and prothrombin fragment F1 + 2 (F1 + 2); anti-FXa activity was tested only in treated thrombophilic pregnant women. F1 + 2 levels progressively increased during pregnancy in both study groups. However, the F1 + 2 increase in women exposed to heparin prophylaxis was significantly lower than that in normal pregnant women in all 3 measurements carried out during gestation (p < 0.05); a statistically significant inverse correlation between F1 + 2 levels and anti-Xa activity (R = -0.8575, p < 0.05) was observed in treated women during pregnancy. Our findings suggest that F1 + 2 in addition to anti-Xa measurement could be used to adjust LWMH prophylaxis, at least in high-risk pregnant women.

VL - 137 IS - 4 U1 - http://www.ncbi.nlm.nih.gov/pubmed/28478442?dopt=Abstract ER - TY - JOUR T1 - Pitfalls and promises in FTIR spectromicroscopy analyses to monitor iron-mediated DNA damage in sperm. JF - Reprod Toxicol Y1 - 2016 A1 - Pascolo, Lorella A1 - Bedolla, Diana E A1 - Vaccari, Lisa A1 - Venturin, Irene A1 - Cammisuli, Francesca A1 - Gianoncelli, Alessandra A1 - Mitri, Elisa A1 - Giolo, Elena A1 - Luppi, Stefania A1 - Martinelli, Monica A1 - Zweyer, Marina A1 - Ricci, Giuseppe AB -

Many drugs, chemicals, and environmental factors can impair sperm functionality by inducing DNA damage, one of the important causes of reduced fertility potential. The use of vibrational spectromicroscopy represents a promising approach for monitoring DNA integrity in sperm, although some limitations exist, depending from the experimental conditions. Here, we report that when using FTIR spectromicroscopy to reveal oxidative stress mediated by Fenton's reaction on hydrated sperm samples, DNA damage interpretation is partially compromised by unexpected cell surface precipitates. The precipitates give a broad band in the 1150-1000cm(-1) infrared region, which partially covers one of the signatures of DNA (phosphate stretching bands), and are detected as iron and oxygen containing material when using XRF spectroscopy. On the other hand, the analyses further support the potential of FTIR spectromicroscopy to reveal cellular oxidative damage events such as lipid peroxidation, protein misfolding and aggregations, as well as DNA strain breaks.

VL - 61 U1 - http://www.ncbi.nlm.nih.gov/pubmed/26923261?dopt=Abstract ER - TY - JOUR T1 - Effect of seminal leukocytes on in vitro fertilization and intracytoplasmic sperm injection outcomes. JF - Fertil Steril Y1 - 2015 A1 - Ricci, Giuseppe A1 - Granzotto, Marilena A1 - Luppi, Stefania A1 - Giolo, Elena A1 - Martinelli, Monica A1 - Zito, Gabriella A1 - Borelli, Massimo KW - Adult KW - Female KW - Fertilization in Vitro KW - Flow Cytometry KW - Humans KW - Leukocytes KW - Male KW - Pregnancy KW - Pregnancy Rate KW - Prospective Studies KW - Semen KW - Sperm Injections, Intracytoplasmic AB -

OBJECTIVE: To investigate the influence of seminal leukocytes on conventional IVF and intracytoplasmic sperm injection (ICSI) outcomes, using a flow cytometry method.

DESIGN: Prospective study.

SETTING: Tertiary infertility center and research institute.

PATIENT(S): One hundred sixty-four couples undergoing conventional IVF or ICSI.

INTERVENTION(S): Seminal leukocytes were counted by flow cytometry.

MAIN OUTCOME MEASURE(S): Correlation between seminal leukocytes concentration and reproductive outcomes in IVF and ICSI cycles.

RESULT(S): The median number of oocytes retrieved, the fertilization and cleavage rate, the median number and grade of embryos transferred, the median number of good-quality embryos transferred, and the median percentage of good-quality embryos from total embryos transferred, in leukocytospermic and non-leukocytospermic patients were not statistically different after either IVF or ICSI. Similarly, there were no significant differences between the two groups for implantation rate and clinical pregnancy rate. Multivariate logistic regression analysis showed that the reproductive outcomes were not influenced by adjustment for female age, infertility diagnosis, number of previous attempts, treatment protocol (GnRH agonist or antagonist), assisted reproduction procedure (IVF or ICSI), and leukocytospermia. By profiling the proper Poisson regression models, no leukocytospermia cut-off value was able to identify the subjects at risk for oocyte fertilization or embryo cleavage failure.

CONCLUSION(S): Using a flow cytometry method, we demonstrated that leukocytospermia does not significantly influence IVF or ICSI outcomes. The same results were obtained by using lower or higher cut-off values for leukocytospermia (from 0.2 to 2 × 10(6)/mL).

VL - 104 IS - 1 U1 - http://www.ncbi.nlm.nih.gov/pubmed/25936234?dopt=Abstract ER - TY - JOUR T1 - Ultrastructural Morphology of Sperm from Human Globozoospermia. JF - Biomed Res Int Y1 - 2015 A1 - Ricci, Giuseppe A1 - Andolfi, Laura A1 - Zabucchi, Giuliano A1 - Luppi, Stefania A1 - Boscolo, Rita A1 - Martinelli, Monica A1 - Zweyer, Marina A1 - Trevisan, Elisa AB -

Globozoospermia is a rare disorder characterized by the presence of sperm with round head, lacking acrosome. Coiling tail around the nucleus has been reported since early human studies, but no specific significance has conferred it. By contrast, studies on animal models suggest that coiling tail around the nucleus could represent a crucial step of defective spermatogenesis, resulting in round-headed sperm. No observations, so far, support the transfer of this hypothesis to human globozoospermia. The purpose of this work was to compare ultrastructural morphology of human and mouse model globozoospermic sperm. Sperm have been investigated by using scanning and transmission electron microscopy. The images that we obtained show significant similarities to those described in GOPC knockout mice, an animal model of globozoospermia. By using this model as reference, we were able to identify the probable steps of the tail coiling process in human globozoospermia. Although we have no evidence that there is the same pathophysiology in man and knocked-out mouse, the similarities between these ultrastructural observations in human and those in the experimental model are very suggestive. This is the first demonstration of the existence of relevant morphological homologies between the tail coiling in animal model and human globozoospermia.

VL - 2015 U1 - http://www.ncbi.nlm.nih.gov/pubmed/26436098?dopt=Abstract ER - TY - JOUR T1 - Medical treatments for endometriosis-associated pelvic pain. JF - Biomed Res Int Y1 - 2014 A1 - Zito, Gabriella A1 - Luppi, Stefania A1 - Giolo, Elena A1 - Martinelli, Monica A1 - Venturin, Irene A1 - Di Lorenzo, Giovanni A1 - Ricci, Giuseppe KW - Endometriosis KW - Female KW - Gonadotropin-Releasing Hormone KW - Histone Deacetylase Inhibitors KW - Hormone Antagonists KW - Humans KW - Pain Management KW - Pelvic Pain KW - Progestins AB -

The main sequelae of endometriosis are represented by infertility and chronic pelvic pain. Chronic pelvic pain causes disability and distress with a very high economic impact. In the last decades, an impressive amount of pharmacological agents have been tested for the treatment of endometriosis-associated pelvic pain. However, only a few of these have been introduced into clinical practice. Following the results of the controlled studies available, to date, the first-line treatment for endometriosis associated pain is still represented by oral contraceptives used continuously. Progestins represent an acceptable alternative. In women with rectovaginal lesions or colorectal endometriosis, norethisterone acetate at low dosage should be preferred. GnRH analogues may be used as second-line treatment, but significant side effects should be taken into account. Nonsteroidal anti-inflammatory drugs are widely used, but there is inconclusive evidence for their efficacy in relieving endometriosis-associated pelvic pain. Other agents such as GnRH antagonist, aromatase inhibitors, immunomodulators, selective progesterone receptor modulators, and histone deacetylase inhibitors seem to be very promising, but there is not enough evidence to support their introduction into routine clinical practice. Some other agents, such as peroxisome proliferator activated receptors-γ ligands, antiangiogenic agents, and melatonin have been proven to be efficacious in animal studies, but they have not yet been tested in clinical studies.

VL - 2014 U1 - http://www.ncbi.nlm.nih.gov/pubmed/25165691?dopt=Abstract ER - TY - JOUR T1 - Soluble TRAIL is present at high concentrations in seminal plasma and promotes spermatozoa survival. JF - Reproduction Y1 - 2014 A1 - Zauli, Giorgio A1 - Celeghini, Claudio A1 - Monasta, Lorenzo A1 - Martinelli, Monica A1 - Luppi, Stefania A1 - Gonelli, Arianna A1 - Grill, Vittorio A1 - Ricci, Giuseppe A1 - Secchiero, Paola KW - Adult KW - Apoptosis KW - Enzyme-Linked Immunosorbent Assay KW - Flow Cytometry KW - Humans KW - Infertility, Male KW - Male KW - Receptors, TNF-Related Apoptosis-Inducing Ligand KW - Semen KW - Sperm Capacitation KW - Sperm Motility KW - Spermatozoa KW - TNF-Related Apoptosis-Inducing Ligand AB -

The expression of tumor necrosis factor-related apoptosis-inducing ligand (TRAIL(TNFSF10)) and of its receptors (TRAILR1, TRAILR2, TRAILR3, and TRAILR4) have been documented in testis, but the presence of soluble TRAIL in seminal fluid, as well as the potential physiopathological role of the TRAIL/TRAILR system in spermatozoa, has not been previously investigated. Male donors (n=123) among couples presenting for infertility evaluation were consecutively enrolled in this study. The presence of soluble TRAIL was analyzed in seminal samples by ELISA, while the surface expression of TRAIL receptors was investigated by flow cytometry. High levels of soluble TRAIL were detected in seminal plasma (median, 11 621 pg/ml and mean±s.d., 13 371±8367 pg/ml) and flow cytometric analysis revealed a variable expression of TRAIL receptors in the sperm cellular fraction among different subjects. In addition, the effect of physiologically relevant concentrations of recombinant TRAIL was investigated on survival and motility of spermatozoa. Of interest, the in vitro exposure of capacitated spermatozoa to recombinant TRAIL (10 ng/ml) significantly preserved their overall survival. Therefore, the present study demonstrates for the first time the presence of elevated levels of the anti-inflammatory cytokine TRAIL in seminal fluids. Moreover, the demonstration that recombinant TRAIL promotes spermatozoa survival after capacitation suggests potential therapeutic implications.

VL - 148 IS - 2 U1 - http://www.ncbi.nlm.nih.gov/pubmed/24825910?dopt=Abstract ER - TY - JOUR T1 - Association between the JC polyomavirus infection and male infertility. JF - PLoS One Y1 - 2012 A1 - Comar, Manola A1 - Zanotta, Nunzia A1 - Croci, Eleonora A1 - Murru, Immacolata A1 - Marci, Roberto A1 - Pancaldi, Cecilia A1 - Dolcet, Ornella A1 - Luppi, Stefania A1 - Martinelli, Monica A1 - Giolo, Elena A1 - Ricci, Giuseppe A1 - Tognon, Mauro KW - Adult KW - Amino Acid Sequence KW - Amino Acid Substitution KW - Base Sequence KW - BK Virus KW - Capsid Proteins KW - DNA, Viral KW - Humans KW - Infertility, Male KW - JC Virus KW - Male KW - Molecular Sequence Data KW - Polyomavirus Infections KW - Semen KW - Sequence Analysis, DNA KW - Tumor Virus Infections AB -

In recent years the incidence of male infertility has increased. Many risk factors have been taken into consideration, including viral infections. Investigations into viral agents and male infertility have mainly been focused on human papillomaviruses, while no reports have been published on polyomaviruses and male infertility. The aim of this study was to verify whether JC virus and BK virus are associated with male infertility. Matched semen and urine samples from 106 infertile males and 100 fertile males, as controls, were analyzed. Specific PCR analyses were carried out to detect and quantify large T (Tag) coding sequences of JCV and BKV. DNA sequencing, carried out in Tag JCV-positive samples, was addressed to viral protein 1 (VP1) coding sequences. The prevalence of JCV Tag sequences in semen and urine samples from infertile males was 34% (72/212), whereas the BKV prevalence was 0.94% (2/212). Specifically, JCV Tag sequences were detected in 24.5% (26/106) of semen and 43.4% (46/106) of urine samples from infertile men. In semen and urine samples from controls the prevalence was 11% and 28%, respectively. A statistically significant difference (p<0.05) in JCV prevalence was disclosed in semen and urine samples of cases vs. controls. A higher JC viral DNA load was detected in samples from infertile males than in controls. In samples from infertile males the JC virus type 2 strain, subtype 2b, was more prevalent than ubiquitous type 1. JCV type 2 strain infection has been found to be associated with male infertility. These data suggest that the JC virus should be taken into consideration as an infectious agent which is responsible for male infertility.

VL - 7 IS - 8 U1 - http://www.ncbi.nlm.nih.gov/pubmed/22912758?dopt=Abstract ER -